New estrane derivatives

ABSTRACT

NEW 14A, 17A-ALKYLIDENEDIOXY AND 14A, 17A-BENZYLIDENEDIOXYESTRANE DERIVATIVES ARE DISCLOSED WHICH POSSESS OESTROGENIC AND HYPOCHOLESTEROLENIC ACTIVITIES.

United States Patent 3,585,192 NEW ESTRANE DERIVATIVES Arthur F. Marx and Hermanus J. Kooreman, Delt't, Netherlands, assignors to Koninklijke Nederlandsche Gist-En Spiritusfabriek N.V., Delft, Providence of South Holland and Kingdom of the Netherlands No Drawing. Filed Sept. 9, 1969, Ser. No. 856,483 Claims priority, application Great Britain, Sept. 11, 1969, 43,295/69 Int. Cl. C07c 173/00 Cl. 260-23955 11 Claims ABSTRACT OF THE DISCLOSURE New l4a,17a-alkylidenedioxy and 14a,17a-benzylidenedioxyestrane derivatives are disclosed which possess oestrogenic and hypocholesterolenic activities.

BACKGROUND OF THE INVENTION This invention relates to new therapeutically useful steroids of the estrane series, to processes for their preparation and to pharmaceutical compositions containing them.

SUMMARY OF THE INVENTION New 14a,17a-methylenedioxy-estrane derivatives have been discovered of the formula DESCRIPTION OF PREFERRED EMBODIMENTS The 14a,17a-methylenedioxy-estrane derivatives of Formula 1 may be prepared by methods known per se for the preparation of analogous compounds. By the term methods known per se is meant methods heretofore used or described in the chemical literature.

According to a feature of the invention the estrane derivatives of Formula 1, wherein R represents an acetoxy group are prepared by reacting 3,14a,17u-trihydroxy-19- nor-1,3,5()-pregnatriene-20-one with an aldehyde of the formula RCHO, wherein R is as hereinbefore defined, followed by converting the thus obtained 14a,17amethylenedioxy derivative of the formula Patented June 15, 1971 wherein R is as hereinbefore defined, to the corresponding 17/3-acetoxy-estrane derivative in manner known per se.

Preferably the reaction of 3,1411,l7a-trihydroxy-l9-nor- 1,3,5(10)-pregnatriene-20-one with an aldehyde of the formula RCHO is carried out at room temperature in the presence of a strong acid as catalyst and, if desired, in an inert organic medium. Preferred catalysts are perchloric acid, p-toluenesulphonic acid, sulphuric acid and hydrochloric acid. Suitable solvents are halogenated alkanes, methyl acetate, ethyl acetate, dioxane, tetrahydrofuran, benzene and dimethylformamide.

The conversion of a 14a,17a-methylenedioxy derivative of Formula 2 to the corresponding 17fl-acetoXy-estrane derivative of Formula 1, wherein R and R; are as hereinbefore defined and R represents an acetoxy group, is preferably carried out in four succeeding reaction steps.

(a) Protection of the 3-hydroxyl group by etherification; for example, a compound of Formula 2 may be reacted with isobutylene, thereby obtaining the corresponding S-tert. butyloxy derivative. The reaction is preferably carried out in an inert organic medium, such as methylene chloride and in the presence of a small amount of a strong acid, such as hydrochloric acid.

(b) A compound of Formula 2, wherein the 3-hydroxyl group has thus been protected, is then converted with hydroxylamine to the corresponding 20-oximino derivative of the formula wherein R and R are as hereinbefore defined. The reaction is preferably carried out in aqueous alkali hydroxide solution.

(c) A 20-oximino compound of Formula 3, is then subjected to a Beckmann rearrangement, for example with phosphorus oxychloride in pyridine at a temperature somewhat below room temperature, for instance, 5-10 C. A 17fl-acetylamino-estrane derivative of the formula wherein R and R are as hereinbefore defined is thereby obtained.

(d) A 17fl-acetylamino-estraine derivative of Formula 4, is converted with a nitrosating agent, for example nitrosyl chloride, to the corresponding 17fl-acetoxy-estrans derivative of Formula 1, wherein R and R are as hereinbefore defined and R represents an acetoxy group. The reaction is preferably carried out in acetic acid and/ or acetic anhydride at a temperature somewhat below room temperature, for instance 5 C.

All the compounds of Formulas 2, 3, and 4 wherein R and R are as hereinbefore defined, are new compounds particularly useful in a preferred procedure for obtained comounds of Formula 1 and as such form a feature of the invention.

The starting material in the procedure described above, 14a,17a trihydroxy 19 nor 1,3,5(10) regnatriene- 20-one, is also a new compound. It can be prepared in a sequence of five reaction steps, using the known compound 14a,l7a-dihydroxyprogesterone as starting material.

(a) First, 14a,l7a-dihydroxyprogesterone may be microbiologically hydroxylated in the ll-position, for example with Cunninghamella blakesleeana, Curvularz'a lunata, or with Aspergillus ochraceus, thus obtaining 1 1,1 40:, 17a-trihydroxyprogesterone.

(b) This compound is then dehydrated to give the corresponding 9(l1)-dehydro derivative; for example, 11a,14a,l7a-trihydroxyprogesterone may be acylated, for example with methane sulphonyl chloride to 1la,l4oz,17atrihydroxyprogesterone ll-methane sulphonate. The acyl derivative, dissolved in a suitable organic solvent, such as dimethylformamide, can then be heated with lithium chloride to give l4a,l7a-dihydroxy-9(1l)-dehydroprogesterone. The llfi-isomer, 11p,14a,17a-trihydroxyprogesterone is converted with N-bromo-acetamide to the corresponding 9(ll)-dehydro derivative. Preferably, this reaction is carried out at room temperature, in a suitable organic medium, such as pyridine.

(c) This compound may be dehydrogenated in the 1,2-position, for example with selenium oxide or 2,3-dichloro-4,5-dicyanobenzoquinone, or by fermentation with a suitable microorganism, such as Corynebacterium simplex, to give 14a,l7a-dihydroxy-1,9(11)-bisdehydroprogesterone, which can also be indicated as 14a,17a-dihydroxy-1,4,9( 1l)-pregnatriene-3 ,20-dione.

(d) The compound thus obtained is then subjected to an aromatization reaction; this can be carried out, for example, by heating the compound in pyridine with zinc dust to give 3,1411,l7a-trihydroxy-19-nor-1,3,5(10)-pregnatetraene-ZO-one.

(e) The 9(11)-double bond is then reduced again, for example catalytically, using platinum oxide as catalyst. There is obtained a mixture of the 911- and the 9,8-isomer, 3, 14a,17a-trihydroxy-19-nor-1,3,5 (l)-pregnatriene 20- one and 3,14a,17atrihydroxy-l9-nor-9 3-l,3,5(10)-pregnatriene-ZO-one respectively, the hydrogen atom in the 9a-position not being indicated as is conventional, which isomers can be separated by crystallization or column chromatography.

Each of these isomers is then reacted further with an aldehyde of the formula RCHO, as is described above, to give respectively the 9aor the 9,3-isomer of a 17fi-acetoxy estrane derivative of Formula 1, wherein R and R are as hereinbefore defined and R represents an acetoxy group.

All the compounds obtained according to the above described processes, except 1113,l4a,l7a-trihydroxyprogesterone, are new.

According to a modification of the procedure described above the l4a,17a-methylenedioxy derivatives of Formula 2, which are intermediates in the process of preparing the compounds of Formula 1, wherein R represents anacetoxy group, are prepared by first converting 14a,l7a-dihydroxyprogesterone to a 14a,l7a-methylenedioxy derivative of the formula wherein R is as hereinbefore defined, and then converting a compound thus obtained to a 3-hydroxy-19-nor-1,3,5- (10)-pregnatriene derivative of Formula 2.

The various processes used to carry out this modified procedure are quite similar to those described hereinbefore. However, upon reduction of the 9(ll)-double bond a mixture of the sand the 9B-isomer is not obtained, as is the case in the first described procedure, only one of the two possible isomers is formed, the nature of the isomer depending on the way in which the reduction is carried out. When the reduction is carried out catalytically, for instance with platinum oxide as catalyst, the 9fi-isomer of a compound of Formula 2 is obtained exclusively, whereas when the reduction of the 9(11)- double bond is carried out with lithium in liquid ammonia only the 9a-isomer is formed.

The compounds of Formula 5 are disclosed in our copending U.S. patent application Ser. No. 712,600, which application is incorporated herein by reference.

According to another feature of the invention, the estrans derivatives of Formula 1, wherein R represents a halogen atom and R and R are as hereinbefore defined, are prepared by reacting a 3-alkoxy-l4a,17a-dihydroxy- 1,3,5 (l0)-estratriene-17,3-carboxylic acid of the formula coon wherein R is as herebefore defined, with an aldehyde of the formula RCHO, wherein R is as hereinbefore defined; the thus obtained l4a,l7a-methylenedioxy derivative of the formula coon ----o (\I i OCHR is then converted to the corresponding 17fi-halo derivative in manner known per se.

The reaction of a 3-a1koxy-14a,l7a-dihydroxy-1,3,5 (10)-estratriene-17B-carboxylic acid of Formula 6 with an aldehyde of the formula RCHO can be carried out in the same manner as described above for the corresponding reaction with 3,l4a,l7a-trihydroxy-19-nor-1,3,5(10)-pregnatriene-20-one.

A compound of Formula 7 is converted to the corresponding l7/3-halo derivative by means of a modified Hunsdiecker reaction (J. Org. Chem. 1961, 26, 280). The reaction is carried out in the presence of lead tetraacetate and a suitable organic solvent medium, such as benzene or carbon tetrachloride. Chlorine can be provided by, for example, lithium chloride; iodine is preferably added as such to the reaction mixture. In the preparation of the l7B-iodoestrane derivatives the reaction mixture is preferably illuminated with a tungsten lamp. The compounds of Formula 6, which are used as startlIlg material in the above preferred procedure, are new compounds. A 3-alkoxy-14a,17a-dihydroxy-1,3,5(10)- estratiene-l7p-carboxylic acid of Formula 6 may be prepared from the known compound l4a-hydroxyhydrocartisone 21-acetate in a manner known per se.

First, a 9(11)-double bond is introduced in l4a-hydroxyhydrocortisone 21-acetate by reaction with N-bromoacetamide in pyridine; then a 1,2-double bond is introduced microbiologically, for example with Corynebacteriam: Simplex, to give 14a.,17a,21- trihydroxy-1,4,9(11)- pregnatriene-3,20-dione 21-acetate. The compound thus obtained is subjected to an aromatization reaction, for example with zinc dust in pyridine by heating under reflux.

There is obtained 3,14a-17u-21-tetrahydroxy-19-nor- 1,3,5(10),9(11)-pregnatetraene-20-one 21-acetate which is converted, after alcoholysis of the acetyl group with for example sodium methanolate in methanol, with trimethyl orthoacetate to 3,14a,l7a-2l-tetrahydroxy-l9-nor-1,3,5 10) ,9 1 1 -pregnatetraene-20-one 17,21-methylorthoacetate. This reaction is preferably carried out at room temperature and under reduced pressure in a suitable organic solvent, such as dioxane, in the presence of a strong acid, such as p-toluenesulphonic acid. After having protected the 17- and 21-hydroxyl groups in this manner, the 3-hydroxyl group is converted to a 3-alkoxy-group, for example With dialkyl sulphate, preferably in a medium consisting of an inert organic solvent, such as chloroform, and an aqueous alkali hydroxide solution; the reaction can be carried out at room temperature. The 17,21-methyl-orthoacetate group is then hydrolyzed again, for example by means of a strong acid, such as hydrochloric acid and is then converted to a 17,21-dihydroxy group.

The 3-alkoxy-14a,17a,21-trihydroxy-19-nor-1,3,5 10) 9(11)-pregnatetraene-20-one thus obtained is oxidized to the corresponding 17/3-carboxylic acid. A suitable oxidation agent is for example periodic acid (H and sodium bismuthate; the reaction can be carried out at room temperature in a suitable organic solvent medium, such as tetrahydrofuran. A 3-alkoxy-14u,17u-dihydroxy-1,3,5 (10),9(11)-estratetraene-17fl-carboxylic acid is obtained which is reduced, for example catalytically, using platinum oxide as catalyst, to a 3-alkoxy-14a,17u-dihydroxy- 1,3,5 (10)-estratriene-17,8-carboxylic acid of Formula 6. A compound of this formula can then be used in the procedure described above to prepare a compound of Formula 1, wherein R represents a halogen atom.

All the compounds described in the above preferred procedure are new compounds particularly useful in the preparation of compounds of Formula 1.

EXAMPLE I (a) 5.5 l. of nutrient medium consisting of 0.5% of glucose and 0.5% of corn steep liquor were inoculated with 275 ml. of shake culture of Aspergillus ochraceus. The microorganism was grown at 26 C. with vigorous stirring and aeration. After 24 hours a solution of 1.4 g. of 14a,17u-dihydroxy-progesterone in ml. of dimethylformamide was added. After 72 hours, the conversion being complete, the culture broth was filtered and the filtrate extracted three times with 1 l. of methyl isobutyl ketone. The extract was concentrated under reduced pressure and the residue crystallized from methanol; yield: 0.4 g. of 11a,14 x,17a-trihydroxyprogesterone.

Melting point: 232-234.5 C. I.R. (in CHCl v ;=3608, 3485, 1711, 1665, 1612,

and 1350 cmf (b) To a stirred suspension of 5 g. of lloz,14ot,17octrihydroxyprogesterone in 50 ml. of pyridine, 1.2 ml. of methane sulphonylchloride was added dropwise. The reaction mixture was kept at room temperature for 90 minutes and then poured into 750 ml. of water, thereby obtaining 5.6 g. of 11a,14u,17u-trihydroxyprogesterone ll-methane sulphonate.

Melting point: 160161 C. LR. (in CHCl v '=36()0, 3505, 1709, 1665, 1610, 1350, 1333, 1170, 921 and 900 cmr (c) A solution of 5.5 g. of 11a,14a,17a-trihydroxyprogesterone l'l-methane sulphonate and 5.5 g. of lithium chloride in 55 ml. of dimethylformamide was kept at 100 C. for minutes. To the cooled reaction mixture 20 ml. of water were added, yielding 3.5 g. of 14,17a-dihydroxy-9 1 1 -dehydroprogesterone.

Melting point: 235-241 C. N.M.R. (in CDClg-l-some DMSO-d 6:0.58, 1.34, 2.15,

about 4.0, 5.58 and 5.68 p.p.m. Molecular ion peak in mass spectrum:

Calculated for C H O 344. Found: 344.

(d) A mixture of 10 g. of 14a,17a-dihydroxy-9(11)- dehydroprogesterone, 6.6 g. of selenium oxide, 2 ml. of pyridine and 500 ml. of tert. butanol was refluxed for 17 hours. The precipitated selenium was filtered off, the filtrate diluted with 2.5 l. of methyl isobutyl ketone and washed with N sodium hydroxide solution, 0.5 N sulfuric acid and water. The organic fraction was concentrated and the brown, crystalline residue triturated with 50 ml. of methanol yielding 2.0 g. of nearly white 1404,17oc-dihydroxy-1,9 1 1 )-bisdehydroprogesterone.

Melting point: 268-272 C. IR. (in CHCl v =3605, 3560, 3475, 1710, 1662,

1622, 1603 and 1350 cm.-

(e) A mixture of 1.0 g. of 14a,17ot-dihydroxy-1,9(11)- bisdehydroprogesterone, 30 ml. of pyridine containing 0.5 ml. of water, and 20 g. of zinc dust was heated under reflux with stirring for 210 min. After cooling, the zinc was removed by filtration and washed with methyl isobutyl ketone. The filtrate was concentrated and the residue dissolved in 25 ml. of methyl isobutyl ketone and this solution was washed with N sulfuric acid and water. The solvent was evaporated in vacuo and the residue was crystallized from methanol; yield: 0.50 g. of 3,14a,17ot.- trihydroxy 19 nor-1,3,5(10),9(11)-pregnatetraene-20- one.

Melting point: 230235 C.

N.M.R. (in a mixture of CDCl and DMSO-d 8:0.68, 2.23, 6.23 (broad), 6.55, 6.6 and 7.5 (AB spectrum) p.p.m.

(f) A solution of 0.40 g. of 3,14a,17a-trihydroxy-19- nor-1,3,5(10),9(11)-pregnatetraene-20-one in 15 ml. of a mixture (1:1) of methanol and methylene chloride was shaken with 10 mg. of platinum oxide for 3 hours in a hydrogen atmosphere. After removal of the catalyst by filtration and concentration of the filtrate there was obtained a residue which was purified by column chromatography (silica gel, eluent: a solution of 5% acetone in benzene): yield: mg. of an a-polar and mg. of a polar compound. The a-polar compound proved to be 3,14a,17a-trihydroxy-l9-nor-9y9-1,3,5 (10) pregnatriene- 20-one.

Melting point: 167-169 C.

N.M.R. (in a mixture of CDCl and DMSO-d 8:0.83,

2.21, 6.67, 6.7 and 7.2 (AB-spectrum) p.p.m.

The polar compound proved to be 3,14a,17a-trihydroxy- 19-nor-1,3,5 (10)-pregnatriene-20-one (i.e. the 9a-isomer). Melting point: 244248 C.

N.M.R. (in a mixture of CDCl and DMSO-d 6:0.72,

2.23, 6.53, 6.6 and 7.1 (AB-spectrum) p.p.m.

(g) A solution of 1.0 g. of 3,14a,17a-trihydroxy-1-9- nor-1,3,5(10)-pregnatriene-20-one in a mixture of 6 ml. of paraldehyde, 6 ml. of dioxane and 0.05 ml. of 70% perchloric acid was kept at room temperature for 15 minutes. On successive addition of 1.5 ml. of N sodium hydrogen carbonate and 30 ml. of water, 1.0 g. of 3- hydroxy-1 4a,17u-ethylidenedioxy 19 nor-1,3,5(10)- pregnatriene-ZO-one was precipitated.

Melting point: 200-201" C. I.R. (in CHCl v =3605, 1708, 1608, 1580, 1492,

1398, 1349, 1130 and 1107 cm:-

(h) A magnetically stirred suspension of 8.5 g. of 3-hydroxy-14u,-17m-ethylidenedioxy 19 nor-1,3,5(10)- pregnatriene-ZO-one and 100 ml. of methylene chloride in a high pressure reaction vessel was cooled to 20 to 30 C. Isobutylene, dried over calcium chloride, was passed in until the volume of the reaction mixture was about 200 ml. Then 0.8 ml. of concentrated sulphuric acid was added, the reaction vessel closed and the contents stirred at room temperature for 72 hours. The reaction mixture was washed with aqueous sodium hydrogen carbonate solution and water and then concentrated to dryness. The residue was crystallized from methanol; yield: 6.9 g. of 3-tert. butyloxy-14a,17a-ethylidenedioxy- 19-nor-1,3,5 10) pregnatriene-20-one.

Melting point: l32141 C. I.R. (in CHCl ,=1709, 1603, 1570, 1490, 1408,

1359, 1165, 1135 and 1112 cm.-

(i) To a solution of 2.0 g. of hydroxylamine hydrochloride in 16 ml. of a aqueous sodium hydroxide solution was added 0.8 g. of 3-tert.butyloxy-14a,17aethylidenedioxy 19 nor-1,3,5 ()-pregnatriene-20-one. Then, ethanol was added (48 ml.) until a clear solution was obtained. The mixture was refluxed for 10 minutes, diluated with water and cooled; the precipitate yielded 0.6 g. of 3 tert.butyloxy--oximino-14a,Nix-ethylidenedioxy-19-nor-1,3,5(10)-pregnatriene.

Melting point: 180184 C. LR. (in cHc1 ,,,=3490, 1600, 1489, 1406, 1365,

1150, 1135 and 1110 cmr (j) A solution of 10 g. of 3-tert.butyloxy-20-oximino- 1411 171: ethylidenedioxy-19-nor-1,3,5(10)-pregnatriene in 40 ml. of pyridine was treated slowly, while stirred at 5-10 C., with a solution of 20 ml. of phosphorus oxychloride in 60 ml. of pyridine and then stirring was continued for three hours at room temperature. The mixture was poured into ice water, neutralized with 4 N hydrochloric acid, and extracted with chloroform. The extract was Washed with water and dried on magnesium sulphate. Evaporation of the solvent gave an oil, which was chromatographed on alumina (eluent benzene/ ethyl acetate). The oil thus obtained was crystallized from benzene; yield: 6.5 g. of 3tert.butyloxy-14a,Nix-ethylidenedioxy- 17;8-acety1amino-1,3,5 10)-estratriene.

Melting point: 126128 C. LR. (in CHCl ):v =3438, 1691, 1603, 1490, 1406 and 1110 cm.-

(k) A solution of 5.5 g. of 3-tert.butyl0xy-14a,l7aethylidenedioxy 1713 acetylamino-1,3,5(10)-estratriene in 180 ml. of acetic acid, 60 ml. of acetic anhydride and 27 ml. of pyridine was treated with 16 g. of anhydrous potassium acetate. The mixture was cooled to 5 C. and then 50 ml. of nitrosyl chloride in acetic anhydride were added dropwise. Stirring was continued for 45 minutes at a temperature between 4 and 6 C. The reaction mixture was poured into ice water and adjusted with an aqueous 4 N sodium hydroxide solution to pH=5 and then extracted with chloroform. The chloroform layer was washed with an aqueous sodium bicarbonate solution and with water until it was neutralized. The extract was dried on sodium sulphate and the solvent evaporated; the residue was chromatographed on alumina (eluent benzene/acetone). Yield: 1.1 g. of 3-tert.butyloxy-14u, 17m-ethylidenedioxy-17fi-acetoxy-1,3 ,5(10)-estratriene.

Melting point: 132l35 C.

A (in methanol) =271 and 278 nm.; E{ =l7 and 19 LR. (in CHCl ):1/ =1748, 1602, 1560, 1490, 1405,

1382,1368, 1135 and 1110 CH1. 1.

N.M.R. (in CDCl ):6=1.00, 1.33, 1.37 (doublet), 2.07, 5.30 (quadruplet), 6.7, 6.8 and 7.2 (AB-spectrum) p.p.m.

Molecular ion peak in mass spectrum:

Calculated for C H O 428. Found: 428.

8 EXAMPLE II (a) According to the procedure described in Example Ig, 3,14a,17u-trihydroxy-19-nor-1,3,5(10) pregnatriene- 20-one (prepared according to the procedure described :in Example If) was converted with hexanal to 3-hydroxy- :,17a hexylidenedioxy-19-nor-1,3,5(10)-pregnatriene- 20-one, isolated as an oil.

LR. (in CHCl ):1/ =3605, 1710, 1610, 1583, 1350 and 1109 cm.- Molecular ion peak in mass spectrum:

Calculated for C H O 412; Found: 412.

(b) According to the procedure described in Example Ih, 3-hydroxy-14a,17ot-hexylidenedioxy-19-nor-1,3 ,5 10)- pregHatriene-ZO-One was converted to 3-tert.butyloxy-14a, 17a hexylidenedioxy-19-nor-1,3,5(10)-pregnatriene-20- one, isolated as an oil.

LR. (in CHCl W 1710, 1606, 1568, 1492, 1385,

1362, 1350, 1148 and 1110 cm.- Molecular ion peak in mass spectrum:

Calculated for C H O 468. Found: 468.

(c) According to the procedure described in Example Ii, 3 tert.butyloxy-14a,17a-hexylidenedi0xy-19-nor-1,3, 5(10)-pregnatriene-20-one was converted to 3-tert.butyloxy 20 oximino-14a,17a-hexylidenedioxy-19-nor-1,3, 5(10)-pregnatriene, isolated as and oil.

I.R. (in CHCl ):1/ =3595, 1650, 1603, 1565, 1490,

1383,1360 and 1109 cm." Molecular ion peak in mass spectrum:

Calculated for C H O N: 483. Found: 483.

(d) According to the procedure described in Example Ij, 3 tert.butyloxy-ZO-oximino-14a,l7a-hexylidenedioxy- 19 nor 1,3,5(10)-pregnatriene was converted to 3-tert. butyloxy 14a,17a-hexylidenedi0xy-17/i-acetylamino-1,3, 5(l0)-estratriene, isolated as an oil LR. (in CHCl )21 =344(), 1690, 1670 (shoulder), 1602, 1490, 1380, 1359, 1105, 986 and 891 cm.- Molecular ion peak in mass spectrum:

Calculated for C H O N: 483. Found: 483.

(e) According to the porcedure described in Example Ik, 3 tert.butyloxy-l4u,17a-hexylidenedioxy-17B-acetylamino-1,3,5(10)-estratriene was converted to 3-tert.butyloxy 140:,17a hexylidenedioxy-l7B-acetoxy-1,3,5(10)- estratriene, isolated as an oil.

A (in methanol) =270 and 278 nm.; E}Z ,=19 and 17 EXAMPLE III (a) According to the procedure described in Example Ig, 3,14a,17 x-trihydroxy l9 nor-1,3,5 (10)-pregnatriene-20-one (prepared according to the procedure of Example If) was converted 'with benzaldehyde to 3-hydroxy-14u,l7a-benzylidenedioxy 19 nor 1,3,5(10)- pregnatriene-ZO-one.

Meltingpoint: 184-186 C. I.R. (in CHCl v ,,,=3608, 1714, 1610, 1585, 1497,

1353 and 1090 cmr (b) According to the procedure described in Example Ih, 3 hydroxy 140:,170: benzylidenedioxy 19 nor- 1,3,5(10)-pregnatriene 20 one was converted to 3- tert.butyloxy-14a,17u-benzylidenedioxy 19 nor 1,3, 5(10)-pregnatriene-20-one.

Melting point: 139.5-140.5 C. IR. (in CHCl v ,,,,=1712, 1608, 1570, 1495, 1364,

1355, 1150 and 1090 cm.-

(c) According to the procedure described in Example Ii, 3-tert.butyloxy 14a,17a benzylidenedioxy 19 nor- 1,3,5() pregnatriene 20 one was converted to 3-tert.butyloxy 20 oximino-l4a,17a-benzylidenedioxy- 19-nor-1,3,5 10)-pregnatriene.

Melting point: 212-213 C. I.R. (in CHC1 v =3598, 1642, 1605, 1565 and (d) According to the procedure described in Example Ij, 3-tert.butyloxy 20 oximino 14a,17a lbenzylidenedioxy 19 nor 1,3,5 (10)pregnatriene Was converted to 3 tert.butyloxy 140:,17oc benzylidenedioxy-17flacetylamino-1,3,5 (10)-estratriene.

Melting point: 177-179 C. I.R. (in CHCl v =344(), 1700, 1692, 1605, 1492,

1386, 1362 and 1090 cmr (e) According to the procedure described in Example Ik, 3 tert.butyloxy 14oc,17oc benzylidenedioxy-17fiacetylamino 1,3,5 10) estratriene was converted to 3-tert.butyloxy 14a,17a benzylidenedioxy-17fl-acetoxy- 1,3,5 10)-estratriene, isolated as an oil.

k (in methanol)=221 (shoulder), 282 and 278 um. E{",,, =190, 19 and 19 LR. (in CHC1 v =1750, 1603, 1583, 1490, 1382, 1361, 1300, 1161, 1149, 1128, 1060, 1050, 1023, 997, 939 and 895 CIIL 1.

N.M.R. (in CD01 5:1.06, 1.32, 2.06, 6.11, 6.73, 6.8 and 7.25 (AB-spectrum) and 7.3-7.7 (multiplet) p.p.m.

Molecular ion peak in mass spectrum:

Calculated for C H O 490. Found: 490.

EXAMPLE IV (a) A mixture of 5.0 g. of 14a-hydroxyhydrocortisone 21-acetate, 3.0 g. of N-bromo-acetamide and 50 ml. of pyridine was stirred for minutes in a nitrogen atmosphere. Then the reaction mixture was cooled to 10 C.; sulphur dioxide gas was passed through the solvent until the mixture gave a negative reaction on acidified potassium iodide/starch paper. On addition of 200 ml. of water the steroid precipitated; yield: 4.4 g. of l4ot,l7zx,21- trihydroxy-4,9 (1 1)-pregnadiene-3 ,20-dione 21-acetate.

Melting point: 187-190 C. LR. (in CHC1 v =3605, 3555, 3405, 1743, 1729,

1665,1612 and 1373 cmr (b) 10 l. of nutrient medium consisting of 1% yeast extract and 0.5% peptone of pH 6.8 were inoculated with 500 m1. of shake culture of Corynebacterium simplex. The micro-organism was grown at 28 C. with vigorous stirring and strong aeration. After 12 hrs. 2 g. of ballmilled 14a,17ot,21 trihydroxy 4,9(11) pregnadiene- 3,20-dione 21-acetate were added. After 24 hours, the conversion being complete, the fermentation fluid was three times extracted with 10 l. of methyl isobutyl ketone; the extract was concentrated and the residue crystallized from methanol. Yield: 0.5 g. of 14a,17a,21-trihydroxy- 1,4,9 1 1 -pregnatriene-3,20-dione 21-acetate.

Melting point: 196199 C. LR. (in CHCI v =3605, 3560, 3460, 1745, 1730,

1665, 1625, 1608 and 1372 cm.-.

(0) A mixture of 1.0 g. of 14a,17a,21-trihydroxy- 1,4,9(11)-pregnatriene-3,20-dione 21-acetate, 30 ml. of pyridine containing 0.5 ml. of water, and g. of zinc dust was heated under reflux with stirring for 210 minutes. After cooling, the zinc was removed by filtration and washed with methyl isobutyl ketone. The filtrate was concentrated and the residue dissolved in 25 ml. of

10 methyl isobutyl ketone and this solution was washed with N; sulphuric acid and water. The solvent was evaporated in vacuo; the residue was crystallized from methanol. Yield: 0.50 g. of 3,14a,17a,21-tetrahydroxy 19 nor- 1,3,5 10) ,9 1 1 -pregnatetraene-20-one 21-acetate.

Melting point: 212-213 C.

N.M.R. (in a mixture of CDCl and DMSO-d 5:0.71,

2.15, 5.00, 6.25 (multiplet), 6.56, 6.65 and 7.55 (AB- spectrum) p.p.m.

(d) A mixture of 10 g. of 3,14a,17a,21-tetrahydroxy- 19 nor 1,3,5(10),9(11) pregnatetraehe-ZO-one 21- acetate, ml. of methanol and 100 ml. of methylene chloride was treated with 25 ml. of N sodium methanolate in methanol and stirred for 30 minutes in a nitrogen atmosphere at room temperature. After acidification with acetic acid, the reaction mixture was concentrated and the residue dissolved in a mixture of water and methyl isobutyl ketone. The organic fraction was separated and washed with Water. On evaporation of the solvent in vacuo an oily residue was obtained which crystallized from methanol/Water. Yield: 7.0 g. of 3,14a,17a,21-tetrahydroxy 19 nor 1,3,5(10),9(11) pregnatetraene- 20-one.

Melting point: 201205 C. I.R. (in KBr): v =3560, 3380, 3270, 3055, 3021, 1728,

1721, 1628, 1610,1570 and 1492 cmr (e) In a reaction vessel suitable for vacuum distillation a mixture of 10 g. of 3,14u,17a21-tetrahydroxy- 19-nor-1,3,5(10),9(11) pregnatetraene 2O one, 50 ml. of dioxane and 50 ml. of trimethyl orthoacetate was kept at room temperature and treated with five 70 mg. portions of p-toluene sulphonic acid over a period of 45 minutes, the pressure in the apparatus being maintained at about 30 mm. Hg. The reaction mixture was neutralized with pyridine and poured into 800 ml. of water. The precipitate yielded 10.6 g. of 3,140t,170t,21- tetrahydroxy 19 nor 1,3,5 (10),9(11) pregnatetraene-20-one 17,21-methylorthoacetate.

Melting point: 171176 C. LR. (in CHCl v ,,,,=3598, 3535, 1725, 1625, 1603,

1572, 1489 and 1389 cm.-

(f) A mixture of 5 g. of 3,14a,17a,21-tetrahydroxy-19- nor-1,3,5(10),9(11)-pregnatetraene-20-one 17 ,2 l-methylorthoacetate, 100 ml. of chloroform, 100 ml. of an aqueous potassium hydroxide solution and 20 ml. of dimethyl sulphate was stirred for 45 minutes at room temperature. The chloroform layer was separated, washed three times with water and evaporated to dryness. After crystallization from methanol there was obtained 4.2 g. of 3-methoxy- 14a,17a,21-trihydroxy-19 nor 1,3,5(l0),9(11)-pregnatetraene-20-one 17,21 methylorthoacetate.

Melting point: 157-161" C.

N.M.R. (in CDCI ):6=O.73, 1.58, 3.30, 3.78, 4.10 and 4.25 (AB-spectrum), 6.32( broad), 6.63, 6.7 and 7.6 (AB-spectrum) p.p.m.

(g) A mixture of 7.0 g. of 3-methoxy-14a,17a,21-trihydroxy-19-nor-1,3,5(10),9(11)-pregnatetraene 20 one 17,2l-methylorthoacetate, 60 ml. of dimethylformamide and 35 ml. of water, adjusted at pH 3 with N hydrochloric acid, Was stirred at room temperature for minutes. The pH of the reaction mixture was then changed to 7 by addition of a saturated sodium hydrogen carbonate solution. Stirring was continued for another 18 hours at room temperature; the mixture was then diluted with 75 ml. of water. The precipitate yielded 6.3 g. of 3-methoxy- 14a,17a,21-trihydroxy 19 nor-1,3,5(10),9(11)-pregnatetraene-20-one-2l-acetate.

Melting point: 209213 C. I.R. (in CHCI :r/ =3605, 3555, 346-5, 1743, 1730,

1619, 1605, 1568, 1492 and 1040 emf.

(h) According to the procedure described above in (d), 3 methoxy 14a,17a,21 trihydroxy 19 nor-1,3,5(10),

1 l 9(11)-pregnatetraene-20-one ZI-acetate was converted to 3-methoxy-14a,17a,21-trihydroxy 19 nor 1,3,5(), 9(11)-pregnatetraene-20-one.

Melting point: 167-171 C. IR. (in CHCl )2v =36l0, 3555, 3485, 1712, 1620,

1606, 1568 and 1493 cmr (i) To a solution of 1.8 g. of 3-methoxy-14a,17a,21- trihydroxy 19 nor-1,3,5(10),9(11)-pregnatetraene-20- one in a mixture of 50 ml. of tetrahydrofuran and 7 ml. of water, a solution of 1.25 g. of crystallized periodic acid (H IOI in 5 ml. of water was added slowly. After minutes again 0.50 g. of periodic acid in 2 ml. of water was added. The reaction mixture was kept at room temperture for 30 minutes and then treated with a solution of 3.0 g of sodium disulphite in 5 ml. of water. The iodine formed was reduced with sodium thiosulphate. The organic layer was salted out, .was separated and the aqueous layer was extracted with methyl isobutyl ketone and the combined organic fractions were concentrated to dryness. The residue was purified and finally crystallized from methanol/ water. Yield: 1.0 g. of 3 methoxy-14a,17a-dihydroxy- 1,3,5(10),9(11)-estratetraene-17/9-carboxylic acid.

Melting point: 184-188 C. I.R. (in CHCl ):v =360O, 3550, 3375, 1768, 1708,

1618, 1603, 1568 and 1492 cm.-

Melting point: 162-164 C. LR. (in CHCl ):u =361O, 3400, 2840, 1769, 1710,

1608, 1570 and 1495 cm.-

(k) A solution of 2.0 g. of 3-methoxy-14u,17a-dihydroxy-l,3,5(10)-estratriene-17,6-carboxylic acid in a mixture of 12 ml. of paraldehyde, 12 ml. of dioxane and 0.1 ml. of 70% perchloric acid was kept at room temperature for minutes. Successively were added 3.0 ml. of aqueous N sodium hydrogen carbonate solution and 60 ml. of water. The precipitate yielded 2.0 g. of 3-methoxy-14a, 17oz ethylidenedioxy-1,3,5(10)-estratriene-flit-carboxylic acid.

Melting point 171-174 C. LR. (in CHCl ):1 =3435, 1772, 1710, 1609, 1573,

1498, 1402, 1348 and 1109 cmr (l) A mixture of 2.0 g. of 3-methoxy-14u,17a-ethylidenedioxy-1,3,5( 10) estratriene 175 carboxylic acid, 2.8 g. of lead tetra-acetate, 0.23 g. of lithium chloride and 65 ml. of benzene were refluxed in a nitrogen atmosphere for minutes. The cooled mixture was diluted with 150 ml. of methyl isobutyl ketone, washed with water and the solvent evaporated in vacuo. Yield: 0.40 g. of 17/3-chloro- 3 methoxy 14a,17a-ethylidenedioxy 1,3,5 (10)-estratriene, isolated as an oil.

km: (in methanol) =225 (shoulder), 278 and 286 nm.;

LR. (in CHCl v =2860, 1609, 1570, 1493, 1400,

1158, 1135, 1100, 1050, 979 and 860 cm.-

N.M.R. (in CDCl ):6=1.05, 1.39 (doublet), 3.79, 5.28 (quadruplet) 6.66, 6.70 and 7.25 (AB-spectrum) p.p.m.

Molecular ion peak in mass spectrum:

Calculated for C H 0 Cl: 362/364. Found: 362/ EXAMPLE V (a) According to the procedure described in Example IVk, 3-rnethoxy-14a,17a-dihydroxy-1,3,5(10)-estratriene- 12 17fi-carboxylic acid (prepared according to the procedure described in Example IVa-j) was converted with hexanal to 3-methoxy-14a,17u-hexylidenedioxy 1,3,5(10) estratriene-17/3-carboxylic acid.

Melting point: 135-136" C. I.R. (in CHCl ):z/ =3435, 1770, 1710, 1609, 1573,

1498, 1350 and 1109 cm..

(b) According to the procedure described in Example IVl, 3 methoxy-14a,17a-hexylidenedioxy-1,3,5(10)-estratriene-l7B-carb0xylic acid was converted to 17fi-chloro-3- methoxy-14a,17a-hexylidenedi0xy-1,3,5(10) estratriene, isolated in oil.

A in methanol=225 (shoulder), 279 and 287 nm.; E}' =145, 36 and 34 LR. (in CHCl ):1/ =2858, 1608, 1572, 1495, 1155,

1100, 1035, 1015, 900 and 860 cm.- N.M.R. (in CDCl ):6=0.97 (triplet), 1.14, 3.78, 5.08

(triplet), 6.67, 6.7 and 7.3 (AB-spectrum) p.p.m. Molecular ion peak in mass spectrum:

Calculated for C H O Cl: 418/420. Found: 418/ EXAMPLE VI (a) According to the procedure described in Example IVk, 3 methoxy-14a,17a-dihydroxy-1,3,5(10) estratriene-l7fl-carboxylic acid (prepared according to the procedure described in Example IVa-j) was converted with benzaldehyde to 3-methoxy l4a,17cc benzylidenedioxy- 1,3,5 10)-estratriene-17fi-carboxylic acid.

Melting point: l200 C. IR. (in CHCl ):v =3445, 1770, 1710, 1610, 1573,

1495, 1350 and 1095 cmf (b) According to the procedure described in Example W1, 3 methoxy 14a,Not-benzylidenedioxy-1,3,5(10)- estratriene 17B carboxylic acid was converted to 17,6- chloro-3-methoxy-14a,17a benzylidenedioxy 1,3,5 (10)- estratriene. Melting point: 138 C.

A (in methan0l)=226 (shoulder), 279 and 286 nm.; l i m. l36, 41 and 38 LR. (in CHCI 1 =1609, 1570, 1492, 1380, 1152,

1090, 1048, 990, 870 and 858 cmr N.M.R. (in CD01 5:1.11, 3.79, 6.06, 6.68, 6.75 and 7.25 (AB-spectrum) and 7.3-7.7 (multiplet) p.p.m. Molecular ion peak in mass spectrum: Calculated for C H O Cl: 24/426. Found: 424/426.

EXAMPLE VII To a stirred suspension of 4.4 g. of lead tetra-acetate in 75 ml. of refluxing carbon tetrachloride were added 3.7 g. of 3 methoxy 14a,17u ethylidenedioxy 1,3,5(10)- estratriene 17B carboxylic acid (prepared according to the procedure described in Example IVa-k) and then 1.27 g. of iodine. The reaction mixture was illuminated with a tungsten lamp for 2 hours. The cooled reaction mixture was filtered and the filtrate washed with N perchloric acid and water. On evaporation of the solvent there is obtained 0.9 g. of 17,8-iode-3-methoxy-14a-17a-ethylidenedioxy-1, 3,5(10)-estratriene, isolated as an oil.

x (in methan0l)=225 (shoulder), 277 and 286 nm.;

i?m.= 25, 50 and 42 triene 17,8 carboxylic acid (prepared according to the procedure descrlbed in Example Va) was converted to 17,3 iodo 3 methoxy 14ot,170t hexylidenedioxy-1,3, (10)-estratriene, isolated as an oil.

A (in methanol)=225 (shoulder), 278 and 286 nm.; Ei",?,,,-=165, 38 and 33 LR. (in CHCl v =2845 (shoulder), 1608, 1570,

1490, 1380, 1152, 1104, 1030, 950 and 852 cmf N.M.R. (in CD01 5:0.88 (triplet), 1.00, 3.80, 5.10 (triplet), 6.68, 6.75 and 7.3 (AB-spectrum) p.p.m. Highest peak in mass spectrum:

Calculated for C H O I: 510. Found: 364 [mol- Weight-(mol. weight HI+H O.)]

The invention includes within its scope pharmaceutical preparations containing, as the active ingredient, at least one of the thereapeutically active compounds of Formula 1 in association with the pharmaceutically acceptable carrier. The preparations may take any of the forms customarily employed for administration of therapeutically active substances, but the preferred types are those suitable for oral administration, especially tablets, including sustained release tablets, pills and capsules including the substance and those suitable for parenteral administration. The tablets and pills may be formulated in the usual manner with one or more pharmaceutically acceptable diluents or excipients, and include materials of lubricating nature. Capsules made of absorbable material, such as gelatin, may contain the active substance alone or in a mixture, with a solid or liquid diluent. Liquid preparations may be in the form of suspensions, emulsions, syrups or elixirs of the active substance in water or other liquid medium commonly used for mixing orally acceptable pharmaceutical formulations.

The active substance may also be made up in a form suitable for parenteral administration, i.e. as a suspension or emulsion in sterile water or an organic liquid usually employed for injectable preparations, for example a vegetable oil such as corn or olive oil, or a sterile solution in water or 'an organic solvent.

For parenteral administration to adult humans the daily dosage may range from 0.1 to 1.0 g.; a suitable concentration of the active principle in the injectable preparation is 100 mg./ml. A pharmaceutical preparation which can be used for human therapy according to the invention is described in the following.

Example g. of 175 chloro 3 methoxy 14a,17a-hexylidenedioxy-1,3,S(l0)-estratriene (prepared according to the procedure described in Example V) were dissolved in 100 ml. of purified corn oil containing 10 of benzyl alcohol (pharmaceutical grade). The solution was filtered through a sterilizing filter. Sterile 'ampoules of 1 ml. were filled under aseptic conditions with 1 ml. of the solution thus obtained.

What we claim and desire to secure by letters patent is:

1. 140:,170c methylenedioxy estrane derivative of the formula wherein R represents an aliphatic hydrocadbon group having less than 6 carbon atoms or a phenyl group, R represents an acetoxy group or a halogen atom and R represents an alkyl group having less than 5 carbon atoms.

2. 14a,17a methylenedioxy estrane derivative according to claim 1, wherein R represents an acetoxy group and R represents a tert. butyl group.

3. 1411,17 methylenedioxy estrane derivative according to claim 1 wherein said derivative is 3-tert.butyloxy- 14,17a ethylidenedioxy 17,8 acetoxy 1,3,5 (10)- estratriene.

4. 1411,1706 methylenedioxy estrane derivative according to claim 1 wherein said derivative is 3-tert.butyloxyl4a,l7ot hexylidenedioxy 17 3 acetoxy 1,3,5(l0)- estratriene.

5. 1406,1706 methylenedioxy estrane dirivative according to claim 1 wherein said derivative is 3-tert.butyloxy- :,17a benzylidenedioxy 17B acetoxy 1,3,5 (10)- estratriene.

6. 1406,1706 methylenedioxy estrane derivative according to claim 1 wherein said derivative is 1713 chloro 3- methoxy-14a,17a-ethylidenedioxy-l,3,5 (10)-estratriene.

7. 14a,l7ot methylenedioxy estrane derivative according to claim 1 wherein said derivative is 17fl-chloro-3- methoxy 1411,1700 hexylidenedioxy 1,3,5(10)-estratriene.

8. 1411,1711 methylenedioxy estrane derivative according to claim 1 wherein said derivative is 17fi-chloro-3- methoxy 14ot,17a benzylidenedioxy-1,3,5,(10)-estratriene.

9. 1400,170: methylenedioxy estrane derivative according to claim 1 wherein said derivative is iodo 3- methoxy 14 x,l7a ethylidenedioxy 1,3,5 (l0)-estratriene.

10. 140:,1700 methylenedioxy-estrane derivative according to claim 1 wherein said derivative is 17,8 iodo-3- methoxy 1411,17 hexylidenedioxy 1,3,5,(10)-estratriene.

11. A 14ot,l7oc methylenedioxy estrane derivative of the formula References Cited FOREIGN PATENTS 9/ 1968 Netherlands ELBERT L. ROBERTS, Primary Examiner G. LOVE, Assistant Examiner US. Cl. X.R. 

